LOCUS       sequence-295635-        8410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8410)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4134192
FEATURES             Location/Qualifiers
     source          1..8410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1916..1936
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2285..2302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2379..2500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2390..2409)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2444..2463
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2615..2661
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2691..2827
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2872..2943
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3169..3757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3249..3268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4257..5466
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5303..5322
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5470..6069
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5470..5489)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5966..5986
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6096..6144
                     /note="synthetic polyadenylation signal"
     CDS             join(6410..6478,6479..7270)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6628..6647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7283..7419
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7564..7635
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7640..7686)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7704..7752
                     /note="synthetic polyadenylation signal"
     enhancer        7833..8136
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8138..8340
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8291..8311
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac actcgctatg tcgataacaa cttctgtggc cctgatggct accctcttga
       61 gtgcattaaa gaccttctag cacgtgctgg taaagcttca tgcactttgt ccgaacaact
      121 ggactttatt gacactaaga ggggtgtata ctgctgccgt gaacatgagc atgaaattgc
      181 ttggtacacg gaacgttctg aaaagagcta tgaattgcag acaccttttg aaattaaatt
      241 ggcaaagaaa tttgacacct tcaatgggga atgtccaaat tttgtatttc ccttaaattc
      301 cataatcaag actattcaac caagggttga aaagaaaaag cttgatggct ttatgggtag
      361 aattcgatct gtctatccag ttgcgtcacc aaatgaatgc aaccaaatgt gcctttcaac
      421 tctcatgaag tgtgatcatt gtggtgaaac ttcatggcag acgggcgatt ttgttaaagc
      481 cacttgcgaa ttttgtggca ctgagaattt gactaaagaa ggtgccacta cttgtggtta
      541 cttaccccaa aatgctgttg ttaaaattta ttgtccagca tgtcacaatt cagaagtagg
      601 acctgagcat agtcttgccg aataccataa tgaatctggc ttgaaaacca ttcttcgtaa
      661 gggtggtcgc actattgcct ttggaggctg tgtgttctct tatgttggtt gccataacaa
      721 gtgtgcctat tgggttccac gtgctagcgc taacataggt tgtaaccata caggtgttgt
      781 tggagaaggt tccgaaggtc ttaatgacaa ccttcttgaa atactccaaa aagagaaagt
      841 caacatcaat attgttggtg actttaaact taatgaagag atcgccatta ttttggcatc
      901 tttttctgct tccacaagtg cttttgtgga aactgtgaaa ggtttggatt ataaagcatt
      961 caaacaaatt gttgaatcct gtggtaattt taaagttaca aaaggaaaag ctaaaaaagg
     1021 tgcctggaat attggtgaac agaaatcaat actgagtcct ctttatgcat ttgcatcaga
     1081 ggctgctcgt gttgtacgat caattttctc ccgcactctt gaaactgctc aaaattctgt
     1141 gcgtgtttta cagaaggccg ctataacaat actagatgga atttcacagt attcactgag
     1201 actcattgat gctatgatgt tcacatctga tttggctact aacaatctag ttgtaatggc
     1261 ctacattaca ggtggtgttg ttcagttgac ttcgcagtgg ctaactaaca tctttggcac
     1321 tgtttatgaa aaactcaaac ccgtccttga ttggcttgaa gagaagttta aggaaggtgt
     1381 agagtttctt agagacggtt gggaaattgt taaatttatc tcaacctgtg cttgtgaaat
     1441 tgtcggtgga caaattgtca cctgtgcaaa ggaaattaag gagagtgttc agacattctt
     1501 taagcttgta aataaatttt tggctttgtg tgctgactct atcattattg gtggagctaa
     1561 acttaaagcc ttgaatttag gtgaaacatt tgtcacgcac tcaaagggat tgtacagaaa
     1621 gtgtgttaaa tccagagaag aaactggcct actcatgcct ctaaaagccc caaaagaaat
     1681 tatcttctta gagggagaaa cacttcccac agaagtgtta acagaggaag ttgtcttgaa
     1741 aactggtgat ttacaaccat tagaacaacc tactagtgaa gctgttgaag ctccattggt
     1801 tggtacacca gtttgtatta acgggcttat gttgctcgaa atcaaagaca cagaaaagta
     1861 ctgtgccctt gcacctaata tgatggtaac aaacaatacc ttcacactca aaggcgaaaa
     1921 cctttacttt cagtcgatga gtgacaaaat cattcatcta accgatgact cctttgatac
     1981 agacgtgttg aaggccgatg gtgcgatact ggtagatttc tgggctgaat ggtgtggccc
     2041 gtgcaagatg atcgcaccca ttctggacga gatcgccgac gagtatcaag gaaaattaac
     2101 tgtcgctaag cttaatatag atcagaaccc agggacggca cctaaatacg gcatcagggg
     2161 aatacctacc ctcctgctct ttaagaatgg ggaagttgcc gcgacaaaag tgggagccct
     2221 gtctaagggc cagctaaaag agttcttgga cgctaacctg gcaggttcag gctcggggca
     2281 catgcaccat caccatcacc atagcagcgg gctcgtgcca cgcggctaag ctagcttgac
     2341 tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg
     2401 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2461 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2521 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2581 caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa
     2641 cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc
     2701 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     2761 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     2821 tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt
     2881 aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc
     2941 ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt
     3001 ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc
     3061 tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa
     3121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3961 agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc
     4021 ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga
     4081 actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc
     4141 caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta
     4201 catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc
     4261 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac
     4321 gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc
     4381 cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg
     4441 gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag
     4501 tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta
     4561 tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg
     4621 ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg
     4681 gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg
     4741 gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca
     4801 aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg
     4861 ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta
     4921 atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg
     4981 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
     5041 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
     5101 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
     5161 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
     5221 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
     5281 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
     5341 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
     5401 aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg
     5461 ttagacaaca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5521 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5581 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5641 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5701 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5761 ttgagcggtt cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg
     5821 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5881 ggtctgggca gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc
     5941 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     6001 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     6061 ggtgcctagg ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat
     6121 ggatgtgtgt tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat
     6181 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     6241 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     6301 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     6361 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca
     6421 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6481 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     6541 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt
     6601 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6661 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc
     6721 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6781 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa
     6841 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6901 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6961 ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca
     7021 gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     7081 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7141 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7201 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7261 gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg
     7321 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
     7381 cttccatacg tgtcctcctt accagaaatt tatccttaag atcgtttaaa ctcgactctg
     7441 gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg
     7501 tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc
     7561 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
     7621 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     7681 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     7741 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
     7801 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     7861 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     7921 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     7981 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     8041 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     8101 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     8161 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     8221 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     8281 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     8341 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     8401 cagcggccgc
//
